Method for determining long non-coding ribonucleic acid interaction proteins
AssigneeCity University of Hong Kong
InventorsLiang Zhang, Jian Yan, Jingyu Li, Wenkai Yi, Ligang Fan
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AssigneeCity University of Hong Kong
InventorsLiang Zhang, Jian Yan, Jingyu Li, Wenkai Yi, Ligang Fan
Estimating the term…
The present invention provides a novel method for determining a long-chain non-coding ribonucleic acid interaction protein. The present invention provides a fusion protein formed by BASU and dCasRx, a mammalian expression vector for expressing said fusion protein. The method for determining the lncRNA interaction protein according to the present invention comprises: co-transfecting a mammalian expression vector that expresses the fusion protein and a gRNA that specifically targets the target lncRNA into target cells, thereby BASU specifically biotin-labeling effector proteins nearby; isolating the biotinylated proteins by using a streptavidin affinity coupled magnetic bead and then eluting, and digesting by trypsin and quantitatively analyzing by a label-free mass spectrometry. The present invention can highly credibly determine the proteins that interact with lncRNA.
A fusion protein formed by biotin ligase BASU and dCasRx, said fusion protein selected from the group consisting of B. subtilis biotin ligase with C-terminal mutation (BASU) as encoded by SEQ ID NO:
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